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Cervical carcinoma induces NLRP3 inflammasome activation and IL-1ß release in human peripheral blood monocytes affecting patients’ overall survival

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Abstract

Purpose

Our group previously demonstrated that genetic variants in inflammasome genes contribute to protection against the establishment of human papilloma virus (HPV)-associated cervical carcinoma (CC). The objective of this study was to better understand the contribution of inflammasome and its cytokines in the CC microenvironment.

Methods

The inflammasome activation was analyzed in CC tumoral cell lines and healthy donors (HD)’ monocytes in co-culture. In vitro results were then compared to CC patients’ public databases.

Results

CC cells did not produce IL-1ß or IL-18 themselves, however, when in co-culture with HD monocytes, induced IL-1ß release in those leucocytes. Inflammasome activation appears to be partially dependent on the NLRP3 receptor. Public data analysis revealed that IL1B expression is increased in the CC compared to normal uterine cervix, and that patients with high IL1B expression had a shorter overall survival.

Conclusion

CC microenvironment can activate the inflammasome and IL-1ß release in surrounding monocytes, which could be detrimental for CC prognosis.

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Acknowledgements

We thank the Blood Bank Service of Hospital “Oswaldo Cruz'' (São Paulo, SP, Brazil), for the Healthy Donors (HD) recruitment; Prof. Luisa Lina Villa, Prof. Ana Paula Lepique, Dr. Lara Termini and Dr. Rafaella Almeida Lima Nunes for reagents donation and critical discussion of the results.

Funding

This work was supported by the São Paulo Research foundation (FAPESP) (Grant n. 2019/06363-4; 2015/50650-7). F.P.F, E.C.R, V.N.C.L. were recipients of a FAPESP fellowship (2018/04361-1, 2017/10824-1, 2015/17373-0). R.A.G.C., and J.L.S. were recipients of a CAPES Fellowship. A.P. is recipient of a CNPq Fellowhsip.

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Authors and Affiliations

  1. Laboratório de Imunogenética, Departamento de Imunologia, Instituto de Ciências Biomédicas (ICB), Universidade de São Paulo (USP), Av. Prof. Lineu Prestes 1730, Cidade Universitária, São Paulo, SP, Brazil

    Fernanda Pereira Fernandes, Raylane Adrielle Gonçalves Cambui, Jaíne Lima da Silva Soares, Edione Cristina dos Reis, Vinícius Nunes Cordeiro Leal & Alessandra Pontillo

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  1. Fernanda Pereira Fernandes
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Correspondence to Raylane Adrielle Gonçalves Cambui.

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Research involving human participants and/or animals and Informed consent

The human samples were provided by the hemotherapy center "Insituto HOC de Hemoterapia" (Sao Paulo), to which belongs the ethical approval. I will e-mail you the term of consent provided by the hemotherapy center.

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12094_2023_3241_MOESM1_ESM.jpg

Supplementary File 1. Inflammaosme genes’ expression in tumoral and normal cervix. Normalized expression of IL1B (A), IL18 (B), NLRP1 (C), NLRP3 (D), NLRC4 (E), NAIP (F), AIM2 (G), CARD8 (H), MEFV (I), CASP1 (J), CASP4 (K), CASP5 (L), and GSDMD (M) in tumoral and normal cervix. Data are from GEPIA software and referred to 306 patients with cervical cancer and 13 healthy donors. Gene expression is expressed as transcript por million cells (TPM). Student t test was applied to compare gene expression in the two groups. Differences with a p-value < 0.05 are considered statistically significant. (JPEG 338 KB)

12094_2023_3241_MOESM2_ESM.docx

Supplementary File 2. IL-1ß plays an apparently favorable role for the growth of CC. Immortalized CC cell lines C-33A, SIHa and HeLa in a three-dimensional (3D) model were cultured in the presence of recombinant human IL-1ß (rh-IL-1ß) (150 pg / mL or 300 pg / mL) for 24-48- 72 hours and the size of the spheroids were measured. (A) A representative photo of the spheres of the SiHa cell line in the presence or not of recombinant human IL-1ß (rh-IL-1ß). (B-D) Bar graph for the three experimental conditions, control, that is, without the presence of recombinant human IL-1ß (rh-IL-1ß), or with 150 pg / mL or 300 pg / mL of recombinant human IL-1ß (rh-IL-1ß). (DOCX 8473 KB)

12094_2023_3241_MOESM3_ESM.docx

Supplementary File 3. The conditioned medium (CM) was not able to lead to the activation of the inflammsome in the monocytes. 6.5 x 104 monocytes (Mo)/well isolated from healthy donors (n=6) were cultured in the presence of an increasing proportion of 2D conditioned medium (CM) of immortalized CC cell lines (A) HeLa and (B) C33-A for 1, 3, 8 and 24 hours. and IL-1ß release was measured in culture supernatants. (DOCX 46 KB)

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Fernandes, F.P., Cambui, R.A.G., Soares, J.L. et al. Cervical carcinoma induces NLRP3 inflammasome activation and IL-1ß release in human peripheral blood monocytes affecting patients’ overall survival. Clin Transl Oncol 25, 3277–3286 (2023). https://doi.org/10.1007/s12094-023-03241-2

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